Regulation of mitochondrial metabolism by lysine acylation

赖氨酸酰化调节线粒体代谢

基本信息

批准号：
9304197
负责人：
ERIC S GOETZMAN
金额：
$ 39.53万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-06-15 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9304197
关键词：
Acetylation Active Sites Acyl CoA Dehydrogenases Acylation Address Affect Binding Binding Proteins Biological Assay Blood Cardiolipins Carnitine O-Palmitoyltransferase Chronic Disease Citric Acid Cycle Complex Defect Development Diabetes Mellitus Diagnosis Dimensions Disease Electron Transport Electrophoresis Energy Metabolism Ensure Enzymes Etiology Fatty Acids Functional disorder Funding Future Genes Genetic Goals Grant Heart Diseases Hereditary Disease Human In Vitro Inborn Errors of Metabolism Inborn Genetic Diseases Individual Inner mitochondrial membrane Kinetics Knockout Mice Knowledge Lipid Binding Liver Long-Chain-Acyl-CoA Dehydrogenase Lysine Macromolecular Complexes Malignant Neoplasms Manipulative Therapies Mass Spectrum Analysis Measures Membrane Membrane Proteins Metabolic Methods Mitochondria Modification Molecular Models Molecular Weight Mouse Protein Muscle function Mutagenesis Myocardium Obesity Organ Pathway interactions Patients Pharmacotherapy Play Police Post-Translational Protein Processing Process Proteins Recombinants Regulation Research Respiratory Chain Role Sirtuins Site-Directed Mutagenesis Testing Therapeutic Work acyl-CoA dehydrogenase deacylation enzyme activity fatty acid metabolism fatty acid oxidation gene replacement heart function improved liver function membrane assembly mitochondrial dysfunction mitochondrial metabolism molecular modeling mortality mouse model novel novel therapeutics operation protein complex

项目摘要

PROJECT ABSTRACT Fatty acid oxidation (FAO) is a critical energy producing pathway in heart, muscle, and liver, among other organs. Inborn errors in genes of the FAO pathway are associated with dysfunction in these organs and a high rate of mortality. Additionally, disruptions in FAO are seen in polygenic diseases such as obesity, diabetes, and cancer. With advances in mass spectrometry profiling of blood metabolites, FAO defects can be readily diagnosed. However, despite 30 years of intensive study, treatment options for modulating FAO in human patients remain limited and ineffective. Knowledge gaps regarding the regulation of FAO enzymes and the functional organization of the FAO pathway within the greater landscape of mitochondrial energy metabolism have limited the development of new therapies. In the previous funding period of this grant, we established reversible lysine post-translational modifications (acetylation, succinylation) as regulators of FAO. We showed that sirtuin enzymes, which deacylate target lysines and restore them to the native state, are important players in maximizing function of the FAO pathway. In the present proposal we hypothesize that lysine acylation regulates FAO enzyme activity, localization to the inner mitochondrial membrane, and the assembly of higher- order metabolic complexes between FAO proteins and the respiratory chain. In Specific Aim 1, we will employ in vitro methods that we pioneered in the previous funding period to identify sirtuin-targeted lysines on the membrane-associated FAO enzymes carnitine palmitoyltransferase-2 (CPT2), mitochondrial trifunctional protein (TFP), and acyl-CoA dehydrogenase-9 (ACAD9). We will perform mutagenesis studies to determine the functional role of each of the sirtuin-targeted lysine residues. In Specific Aim 2 we will investigate physical and functional interactions between the three mitochondrial sirtuins (SIRT3, SIRT4, and SIRT5) and the inner mitochondrial membrane. We hypothesize that the sirtuins police the inner mitochondrial membrane in order to facilitate assembly and operation of higher-order metabolic complexes such as those formed between FAO and the electron transport chain. Finally, Specific Aim 3 will evaluate the effects of lysine acylation on these higher-order complexes using a combination of mouse models and protein complexes assembled in vitro. Understanding the role of the sirtuins in regulating FAO and metabolic supercomplexes will lay the ground work for developing new therapies that manipulate mitochondrial function in human patients with inborn errors of metabolism, as well as those with chronic diseases such as obesity, diabetes, and cancer.

项目摘要脂肪酸氧化 (FAO) 是心脏、肌肉和肝脏等体内重要的能量产生途径器官。 FAO 途径基因的先天性错误与这些器官的功能障碍和高死亡率。此外，肥胖、糖尿病等多基因疾病也造成了粮农组织的破坏。癌症。随着血液代谢物质谱分析的进步，FAO 缺陷可以很容易地被识别出来。确诊。然而，尽管经过 30 年的深入研究，调节人类 FFA 的治疗方案仍然存在。患者仍然有限且无效。关于粮农组织酶的监管和线粒体能量代谢大格局中FAO途径的功能组织限制了新疗法的开发。在本次赠款的上一个资助期间，我们建立了可逆的赖氨酸翻译后修饰（乙酰化、琥珀酰化）作为FAO的调节剂。我们展示了 Sirtuin 酶可以使目标赖氨酸脱酰并将其恢复到天然状态，是重要的参与者最大限度地发挥粮农组织途径的功能。在本提案中，我们假设赖氨酸酰化调节FAO酶活性、线粒体内膜定位以及高级组装排列FAO蛋白质和呼吸链之间的代谢复合物。在具体目标 1 中，我们将采用我们在上一个资助期首创的体外方法，用于识别沉默调节蛋白（sirtuin）靶向的赖氨酸膜相关的FAO酶肉毒碱棕榈酰转移酶-2 (CPT2)，线粒体三功能蛋白 (TFP) 和酰基辅酶 A 脱氢酶 9 (ACAD9)。我们将进行诱变研究以确定每个针对 Sirtuin 的赖氨酸残基的功能作用。在具体目标 2 中，我们将研究物理以及三种线粒体 Sirtuins（SIRT3、SIRT4 和 SIRT5）与内部结构之间的功能相互作用线粒体膜。我们假设去乙酰化酶负责调控线粒体内膜，以便促进高阶代谢复合体的组装和运行，例如粮农组织之间形成的代谢复合体和电子传输链。最后，具体目标 3 将评估赖氨酸酰化对这些的影响使用小鼠模型和体外组装的蛋白质复合物的组合来构建高阶复合物。了解sirtuins在调节FAO和代谢超级复合物中的作用将为我们奠定基础致力于开发操纵先天性缺陷人类患者线粒体功能的新疗法新陈代谢障碍，以及患有肥胖、糖尿病和癌症等慢性疾病的人。